PRRSV-GP5基因的克隆及其B细胞抗原表位预测与分析

PRRSV-GP5 Gene Cloning and Analysis of B Cell Antigen Epitope

【目的】研究PRRSV-GP5基因B细胞抗原表位性质。【方法】根据Gen Bank上发表猪繁殖与呼吸综合征病毒(PRRSV)ATCC VR-2332的全基因序列,设计并合成引物,采用RT-PCR技术,对阳性病料扩增回收,并将其克隆入p MD18-T载体中,送上海生工测序。应用生物信息学同源模型化的方法建立其PRRSV-GP5蛋白3D结构,并根据生物信息学软件DNAStar预测PRRSV-GP5蛋白的二级结构、表面可及性、抗原指数、亲水性及柔韧性,综合分析预测其B细胞抗原表位。【结果】结果表明,其肽段30-36、50-55、140-142、146-151和196-198可能是其优势B细胞抗原表位区域。【结论】该基因高度保守,PRRSV-GP5蛋白呈现较规则的空间结构,同时该结果将为PRRSV-GP5蛋白体外表达产物的应用和基因工程疫苗的研究提供理论依据。

[Objective] The objective of this experiment was to study the B cell epitopes of PRRSV-GP5 gene. [Method] Primers for RT-PCR amplification of PRRSV isolates from Heilongjiang were designed according to the sequence of ATCC VR-2332 strain porcine reproductive and respiratory syndrome virus published in the GenBank, and sequences were cloned into pMD18-T vector respectively and sequenced in Shanghai. The 3D model of PRRSV-GP5 protein was obtained by homology modeling of bioinfor- matics, and DNAStar was used to predict the secondary structure, surface probability, antigenic index, hydrophilicity and flexibility region. [Result] The peptides of PRRSV-GP5 including 30-36, 50-55, 140-142, 146-151 and 196-198 were larvaceous dominant peptide. [Conclusion]The results indicat- ed that the target gene of PRRSV-GP5 was highly conservative and space structure of PRRSV-GP5 was regular. The results will provide theoretical evidence for application of expression product and engineering vaccine of PRRSV-GP5 protein.